RISK ASSESSMENT RESEARCH FOR THE PD PROJECT

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• RISK ASSESSMENT RESEARCH FOR THE PD PROJECT
• Thomas A. Miller1, David J. Lampe2, Ravi
  Durvasula3
• 1University of California, Riverside, CA
• 2Duquesne University, Pittsburgh, PA
  
• 3University of New Mexico, Albuquerque, NM

Regulatory Communication Workshop 7-9 November
2006 UC Washington DC Center
Thomas.miller_at_ucr.edu
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My name is Tom Miller 1. Professor of
Entomology. Teach Insect Physiology and Insect
Toxicology. 2. Entomologist in the Agricultural
Experiment Station. Do something about plant
pests and disease.
Thomas.miller_at_ucr.edu
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August 25, 2005 Los Angeles Times
Hurricanes in 2004 spread the citrus canker
pathogen, Xanthomonas axonopodis from containment
in the 2 southeastern counties of the state where
a 500 million program had pushed it.
The only defense is containment through
destruction.
We had to destroy the village to save it
Attributed to many different people, including
war correspondent Peter Arnett who supposedly
attributed the quote to an unidentified Army
officer. Used circa 1968, perhaps during the
bloody Tet offensive.
Thomas.miller_at_ucr.edu
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"California is particularly vulnerable to
non-indigenous pests and diseases,"
Daniel Sumner, a UC Davis agricultural economist
and director of the Agricultural Issues Center.

Thomas.miller_at_ucr.edu
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A new pest comes into California every 60 days.
Pierces disease first noted in Anaheim in 1884
Orange County disease.
Glassy-winged sharpshooter (GWSS) arrived 1980s?
Thomas.miller_at_ucr.edu
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Total economic impact of more than 170 million a
year, the Texas wine industry has more than 85
wineries and is the nations fifth-largest wine
producing state."
Pierces Disease The most serious threat to wine
grapes in Texas, California and other growing
areas is Pierces disease, which occurs in all
areas of Texas that do not have severe winters.
Pierces disease, which can wipe out a vineyard.
Currently, there is no preventative or curative
treatment for it. Efforts to reduce the risk of
Pierces disease include controlling the
glassy-winged sharpshooter. Site selection and
chemical control help prevent the pathogens
spread. Although a large research effort is
underway to discover methods for controlling
Pierces disease, it will probably be a long
time, perhaps 10 years or more by some estimates,
before a practical, effective solution is
available.
                                                
                                                  
              
Florida grape varieties are resistant to many
diseases, principally Pierce's disease (PD), a
disease that devastates Vitis vinifera. Soon
molecular genetic research will produce further
improved varieties and possibly enable
traditional wine grape production in our state.
Bryon Biddle Three Oaks Winery, Vernon, FL 20
October 2006
Thomas.miller_at_ucr.edu
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Pierces disease control program annual report to
the legislature 2005
Thomas.miller_at_ucr.edu
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Grape acreage by County in California 2005
back in 1999-2000, people were saying that the
grape growing industry was dead in Temecula and
there will be a 100 loss of vines due to
PD. -- Nick Toscano
Thomas.miller_at_ucr.edu
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Callaway vineyards lost about 100 close
proximity to 120 acres of grapefruit 800 out
of a total of 2500 acres of vines were removed
from Temecula in 2001. Since I initiated and
directed the program, 300 new acres have been
planted. The number of wineries have increased
from 13 to 21 (60 increase). -- Nick Toscano
12 October 2006
Nick Toscano Specialist Extension
Entomologistemail nick.toscano_at_ucr.edu(951)827-
5826(951)827-3725
FISCAL YEAR 2003/04 2004/05
2005/06 REVENUE State (Budget Act) 6,408,000
4,408,000 4,341,000 Federal (USDA)
9,885,525 14,903,606 15,300,000 Board
Assessment 968,000 305,000
343,400 Total Resources 17,261,525 19,616,606
19,984,400 EXPENDITURES Personal Services
3,053,409 3,456,754 3,658,520 Operating
Expenses 2,206,328 3,040,503
2,852,186 County Payments 12,001,788 13,119,349
13,473,694 Total Expenditures 17,261,525
19,616,606 19,984,400
Thomas.miller_at_ucr.edu
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Symbiotic control of PD
2. Design toxins against Xylella.
2004 ()
1. Find a symbiont (blue) cycling with the
pathogen (orange).

3. Disease cycle testing.
2001
Axd
Dave Lampe Duquesne University, Pittsburgh PA
2001
DsRed Alcaligenes (RAxd)
S1 RAxd
Endophyte Candidate Alcaligenes xylosoxidans
Tim Yolo and Arinder Arora, UC Riverside
Carol Lauzon CSUEB Hayward CA
2001-2005
2003 Field testing of RAxd?
Thomas.miller_at_ucr.edu
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• Where is risk assessment done?
• In the laboratory at BL-1.
• In the field.
• Laboratory data poorly predict field results.
• GWSS die in 3 weeks in the lab.
• Grapevines do poorly in greenhouses.
• It may be impossible to simulate citrus next to
  vineyards.

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Field testing for Risk Assessment
We want to test behavior of RAxd in grapevines
(2003).
Okay, but you have to burn the grapevines at the
end of the season.- EPA
Then how will we know if the disease is cured or
not?
We dont make the laws. (National Environmental
Protection Act 1969 FQPA 1996 PPA 1990 TSCA
1976 FIFRA 1972).
The UCR Biosafety Committee approved of working
with RAxd at BL-1 level. Why are we burning
grapevines?
appropriate for working with microorganisms
that are not known to cause disease in healthy
human municipal water-testing laboratories, in
high schools, and in some community colleges
teaching introductory microbiology classes
Thomas.miller_at_ucr.edu
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"The Biosafety level is chosen to be commensurate
with the potential risk.  Rather than ensure
absolute containment of every experiment, which
would be onerous for researchers, containment
safeguards are allowed to be more relaxed when
the risk is small.  It is recognized that BL1 may
not ensure containment all of the time, but that
is acceptable since (for example) damage caused
by release is considered insignificant and the
likelihood of the organism to survive outside the
lab is also considered low." -- Howard
Judelson Professor of Plant Pathology UC
Riverside 23 October 2006
BioSafety ruling incompatible with regulation?
Thomas.miller_at_ucr.edu
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Needle inoculation of grapevines, Napa, CA summer
2003.
Bagged grapevines, Temecula, CA 2004
Thomas.miller_at_ucr.edu
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Movement of Alcaligenes in Host Plants

• Plant type Detected/tested amount found
• Lemon 25/25 3,591,427 cells/2cm
• Orange 25/25 943,305 cells/2cm
• Crepe Myrtle 8/25 884,770
  cells/2cm
• Periwinkle 10/25 304,820
  cells/2cm
• Grapevine 24/25 18,225
  cells/2cm

Cells in 2 cm of plant stem
Movement three inches away in two weeks Lemon
preferred
Thomas.miller_at_ucr.edu
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The GloFish
Normal Zebra fish
Los Angeles Times, front page, 22 November 2003
Thomas.miller_at_ucr.edu
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Regulatory wars
Can we sell DsRed zebra fish as pets? -- 2003
Lets see, okay go ahead, we cant stop you,
there is no law against that. USDA, FDA, EPA,
CDC, NIH
Hey, thats the same transgene as in RAxd. Why
dont you make them burn the fish tanks?
We dont make the laws.
Thomas.miller_at_ucr.edu
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November 2003
Thomas.miller_at_ucr.edu
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http//ucbiotech.org/
Regulatory agencies dont need to worry about the
values question others are already doing that.
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Study what researchers think is important.

• Environmental Impact of Transgenic Alcaligenes
  Used to Control Pierces Disease.
• Miller, UCR Durvasula, Yale Lampe, Duquesne
• September 2004.
• Identification of hazards.
• Fitness alteration
• Transfer to non-target organisms
• Transgene instability
• Assessment of community ecology impacts.
• 1. Creating transgenic Axd strains.
• 1a. Attenuated strains.
• 1b. Cysteine auxotrophic strains
• 2. Measuring the rate of horizontal gene transfer
• 3. Monitoring for HGT in natural environments by
  RT PCR.
• Reducing the likelihood of HGT genetic
  toxin-antidote.
• Testing of ecological impact of transgenic Axd.
• 5.1. Bacterial census of grapevine.

Amount requested 200,000/yr Amount funded
100,000/yr Term 2 years.
Thomas.miller_at_ucr.edu
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4 events in 2004
Pew Foundation 22 Jan 2004
Business Week 3 May 2004
NAS Dec 2004
NAS Dec 2004
likelihood of success of symbiotic control
is limited
science knows too little about how the novel
organisms will behave in the real world.
Regulation of transgenic insects is unprepared.
Regulatory and public discourse centers around
the unnaturalness of genetic transformation.
Thomas.miller_at_ucr.edu
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Pages 109-110 Using paratransgenesis to manage
PD clearly would be along-term strategy, and one
in which the likelihood of success is limited.
Although some progress toward transformation of
GWSS endosymbionts has been made (Lampe and
Miller, 2002), the committee views this as
Category 4 research, in addition to its
scientific uncertainty, there are ecological and
regulatory barriers to success that are at least
as significant as any technical barriers.
NAS Dec 2004
COMMITTEE ON CALIFORNIA AGRICULTURAL
RESEARCH PRIORITIES PIERCES DISEASE ____________
_______________________________________________ JA
N E. LEACH, Chair, Kansas State University,
Manhattan PEDRO BARBOSA, University of Maryland,
College Park MICHAEL J. DAVIS, University of
Florida, Homestead DAVID G. HOEL, Medical
University of South Carolina, Charleston L. JOE
MOFFITT, University of Massachusetts,
Amherst ALISON G. POWER, Cornell University,
Ithaca TERRY L. ROOT, Stanford University,
Stanford JACK SCHULTZ, The Pennsylvania State
University, University Park WILLIAM F. SPLINTER,
University of Nebraska, Lincoln BRIAN J.
STASKAWICZ, University of California,
Berkeley MARIE-ANNE VAN SLUYS, University of São
Paolo, Brazil T. ULF WESTBLOM, Central Texas
Veterans Health Care System, Texas AM
University, Temple Staff KIM WADDELL, Study
Director (through March 2004) ROBIN SCHOEN,
Senior Program Officer MICHAEL KISIELEWSKI,
Research Associate DONNA WILKINSON, Research
Intern PETER ROGERS, Research Intern CINDY
LOCHHEAD, Project Assistant JULIE COFFIN, Project
Assistant
This is exactly what they said about transgenic
pink bollworm.
Thomas.miller_at_ucr.edu
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Frances Oldham Kelsey, Ph.D., M.D., (b. 24 June
1914) is a naturalized American pharmacologist,
most famous as the reviewer for the U.S. Food and
Drug Administration (FDA) who refused to
authorize thalidomide for market because she had
concerns about the drug's safety. Her concerns
proved to be justified when it was proven that
thalidomide caused birth defects. Kelsey's career
intersected with the passage of laws
strengthening the FDA's oversight of
pharmaceuticals.
Frances Kathleen Oldham Kelsey receiving the
President's Award for Distinguished Federal
Civilian Service from President John F. Kennedy,
in 1962
Thomas.miller_at_ucr.edu
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13. Regulatory Issues with Biotechnology (Thomas
A. Miller) 1) Margaret McFall-Ngai, Univ.
Wisconsin, Symbiosis as an emerging field
invited, no reply 2) Angela Douglas Univ.York,
UK, Symbiosis as an emerging field accepted in
another session 3) David Brooks, Oxitec, UK
Transgenic insects invited 4) FDA and tooth
decay, Jeffrey Hillman, Oragenics, FL, USA
accepted, health allowing 5) Shrimp
aquaculture, Ravi Durvasula, Univ. of New Mexico
accepted 6) Chagas disease, Ravi Durvasula,
Univ. of New Mexico. accepted 7) India
Regulatory process, lead by K. P. Gopinathan,
Bangalore, India invited 8) China Regulatory
process, lead by Yongping Huang accepted 9)
Thailand and Laos Regulatory process, Yupa
Hanboonsong accepted 10) USA Regulatory
process, Janet L. Andersen, EPA-BPPD invitation
pending 11) USA Regulatory process, Robyn Rose,
APHIS-BRS invitation pending 12) British
Regulatory process, Angela Douglas accepted 13)
Canadian Regulatory process, Brian Belliveau,
Health Canada recruiting someone else 14) Greek
Regulatory process, Hanneke Drosopoulos
accepted 15) Korean Regulatory process,
Ministry of Environment accepted being
organized 16) French Regulatory process, asked
Rene Feyereisen to suggest someone 17) Morocco
Regulatory process, Malika Bounfour invitation
pending 18) Slovenia Regulatory process, Prof.
Dr. Maja Ravnikar accepted